Anti-inflammatory Effect Of Polyphenols From Physalis Alkekengi Fruit

In this paper,the anti-inflammatory effect of polyphenols from Physalis alkekengi fruit were investigated.The research was tested by below directions:extraction conditions of polyphenols,Effect on cell viability,Inflammatory cytokines,mRNA expressions,protein expressions of polyphenols.1.Response surface methodology was employed to optimize extraction process of polyphenols from Physalis alkekengi fruit,ethanol concentration,solid-liquid ratio,extraction temperature and extraction time as factors,the optimized conditions were as follows:the ethanol concentration was 61.5%,the proportion of solid-liquid was 1:15(g:mL),the extraction temperature was 59 ?,and the extraction time was 102 min.Under the optimal conditions,the maximum extraction amounts of polyphenols was 8.986mg/g,extraction ratio was 0.899%.The purity of polyphenols was 60%after purification.2.The effect of the different concentrations polyphenols(4000,2000,1000,500,250,125,62.5,31.25,15.63,7.81,3.91,1.95?g/mL)on RAW 264.7 macrophages viability was tested by the MTT assay.The result indicate:when the concentration was 0-500 ?g/mL,cell proliferation was not affected,and cell morphology did not changed.A majority of cell was damaged when the polyphenols concentration was exceed 500 ?g/mL,and affected the proliferation of RAW 264.7 macrophages.The cell viability was 50%when the concentration was 1136.35 ?g/mL.3.The secretion of NO,PGE2,IL-1?,IL-6,TNF-a in LPS-stimulated RAW 264.7 macrophages was tested by ELLSA assay,and the macrophages was injected different concentration of polyphenols from Physalis alkekengi fruit.Polyphenols(500?g/mL,250?g/mL,125?g/mL)can highly significant or significant(??P<0.01,P<0.05)inhibition the secretion of NO,PGE2,IL-1?,IL-6,TNF-a in RAW 264.7 macrophages,and have a dependence with concentrations.4.The mRNA expressions of inflammatory cytokines in polyphenols from Physalis alkekengi fruit were detection by RT-PCR.Polyphenols in the testing concentrations can highly significant or significant(??P<0.01,?P<0.05)inhibition the mRNA expressions of inflammatory cytokines(iNOS,PGE2,IL-1?,IL-6,TNF-?)in RAW 264.7 macrophages.Inhibition presented a growth trend along with the increase of concentration,the inhibition was most obvious when the polyphenol concentration is 500?g/mL.5.Western blot was employed to analyse the iNOS protein expression that impacted by polyphenols from Physalis alkekengi fruit.The result indicate:the iNOS protein expression on LPS-stimulated RAW 264.7 macrophages can be inhibited by all concentrations(500p?g/mL,250?g/mL,125?g/mL),The results were highly significant(??P<0.01),and have a dependence with concentrations.